DHA (purity ≥98% CAS: 71939‐50‐9) was bought from Yuanye Bio‐Technology Co., Ltd. (Shanghai, China). Puromycin was purchased from Sigma‐Aldrich Corporation (St. Louis, MO, USA). Hoechst 33342 fluorescence staining kit were purchased from Dojindo Laboratories Co., Ltd. (Kumamoto, Japan). Belnacasan, the protease inhibitor Z‐VAD‐FMK (benzyloxycarbonyl‐ValAla‐Asp (OMe) fluoromethylketone), ferrostatin‐1, erastin, deferoxamine and necrostatin‐1 were acquired from MedChemExpress (Monmouth Junction, NJ, USA). An apoptosis detection kit was got from BD Biosciences (San Jose, CA, USA). Foetal bovine serum (FBS) was obtained from Gibco (Thermo Fisher Scientific, Waltham, MA, USA).
Belnacasan
Manufactured by MedChemExpress
Sourced in United States, China
Belnacasan is a laboratory instrument designed for the in vitro analysis of biological samples. It is a multi-functional device that can perform various analytical tasks, including cell-based assays, enzyme activity measurements, and high-throughput screening. The core function of Belnacasan is to facilitate the study of cellular processes and the evaluation of potential pharmaceutical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Wedelolactone, by MedChemExpress (2 mentions)
Deferoxamine, by MedChemExpress (1 mentions)
Necrostatin 1, by MedChemExpress (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Puromycin, by Merck Group (1 mentions)
Apoptosis detection kit, by BD (1 mentions)
Ferrostatin 1, by MedChemExpress (1 mentions)
Z vad fmk, by MedChemExpress (1 mentions)
Erastin, by MedChemExpress (1 mentions)
Pc12 cells, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Rat pheochromocytoma pc12 cells, by American Type Culture Collection (1 mentions)
Methylene blue, by MedChemExpress (1 mentions)
C57bl 6n mice, by Charles River Laboratories (1 mentions)
Disulfiram, by MedChemExpress (1 mentions)
Collagen type 1, by Corning (1 mentions)
Collagenase 4, by Merck Group (1 mentions)
5 protocols using belnacasan
1
Investigating Ferroptosis Inhibitors in Cells
2
Differentiated PC12 Cells: LPS and ATP Treatment
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Rat pheochromocytoma PC12 cells (American Type Culture Collection, Manassas, VA, USA) were cultured in high-sugar basic Dulbecco’s Modified Eagle’s Medium (DMEM) containing penicillin (1 × 105 U/L) and streptomycin (100 mg/L) at 37°C with 5% CO2. Nerve growth factor (NGF, 50 ng/mL) was added (2.5 mL) to the DMEM medium to induce the differentiation of PC12 cells into sympathetic neuron-like cells. The PC12 cells were differentiated continuously for 7 days and the culture solution was changed every 2 days. When the protrusion lengths of the PC12 cells were more than twice the width of the cell body the cells were considered to be completely differentiated. Serum-free medium was added to the differentiated PC12 cells, and 2 hours later, the cells were treated with different concentrations (0.1, 1, 10, and 50 µg/mL) of lipopolysaccharides (LPS; Sigma-Aldrich, St. Louis, MO, USA) at different time points (0, 12, and 24 hours). This was followed by treatment with 5 mM adenosine triphosphate (ATP) for 0.5 hours. Cells in the control group were cultured under normal conditions (no LPS or ATP). These cells were then subjected to treatment with 40 μM Belnacasan (Beln) or 2.5 μM Wedelolactone (Wede) purchased from MedChemExpress (Shanghai, China). This study was approved by the ethics committee of Yantai Yuhuangding Hospital, Qingdao University.
3
Evaluating Combinatorial Immunotherapies in GBM and Melanoma
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GBM orthotopic model: SB mGSC (5 × 103/mouse) or GL261 (105/mouse) were stereotactically injected into the right striatum of six-to-eight-week-old male C57BL/6N mice (Charles River). Melanoma subcutaneous model: B16F10 (2 × 105) was subcutaneously injected into the right axilla of six-to-eight-week-old male C57BL/6N mice (Charles River). Following the tumor injection, mice were randomly divided into four groups: control, anti-mouse PD-L1 mAb (Bio X Cell, BE0101) only, Methylene Blue or Belnacasan (MedChemExpress, HY-13205) only, and the combined treatment group. Methylene Blue (3 mg/kg body weight) was intraperitoneally injected once every two days for two weeks beginning after 3rd day of tumor injection. Belnacasan (50 mg/kg body weight) was intraperitoneally injected every day for two weeks beginning after 3rd day of tumor injection. Anti-mouse PD-L1 mAb (200 µg/mice) was intraperitoneally injected once every three days for three times beginning after 5th day or 7th day of tumor injection in GBM model or melanoma model, respectively. For the melanoma model, tumor volume was measured every five days, and tumor volume was calculated according to the formula: length × (width)2 × 1/2. 20 days after the tumor injection, tumor-bearing mice were sacrificed, and tumor masses were resected for further analysis.
4
Cellular Apoptosis and Inflammation Assays
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Disulfiram (DSF; HY‐B0240), 5‐bromo‐2′‐deoxyuridine (BrdU; HY‐15910), caspase‐11 inhibitor Wedelolactone (HY‐N0551), and caspase‐1 inhibitor Belnacasan (VX‐765; HY‐13205) were obtained from MedChemExpress. Recombinant mouse IL‐1β (50101‐MNAE) and IL‐18 (50073‐MNCE) were obtained from SinoBiological. Collagen type I (354236) was purchased from Corning. Collagenase IV (C5138) was obtained from Sigma‐Aldrich.
5
NLRP3 Inflammasome Modulation in OCHSC
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On 6 DIV, one hour prior to co-culturing PBMCs with OCHSCs, regular OCHSC medium was replaced with serum-free medium, which contains all components of the regular medium except for heat inactivated horse serum. The selective Caspase-1 inhibitor, VX-765 (or Belnacasan, MedChemExpress, stock concentration 196.4 mM dissolved in DMSO) was added to serum-free medium at a nal concentration of 100 µM, and 0.055% nal DMSO volume. Concentration of VX-765 was selected based on an earlier study, which optimized the VX-765 dosage needed to inhibit NLRP3 in ammasome activation in organotypic hippocampal slice cultures (25) . For control experiments, only the vehicle, i.e. DMSO, was added to the medium at the same volume. PBMC-OCHSC co-cultures were incubated with VX-765 overnight, and utilized for electrophysiology and immunohistochemistry experiments the next day, i.e. 7 DIV.
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